Thursday, 23 July 2015

Hepatocyte Cell Line: HepaRG

BioPredic have an exclusive license to manufacture the hepatocyte cell line: HepaRG.

HepaRG is a unique and well established hepatic cell system able to produce early hepatic progenitor cells as well as completely mature human hepatocytes.  It is an alternative to fresh hepatocytesm providing a one-stop hepatic in-vitro model for:

In-vitro ADME applications:

Inhibition assays
Drug metabolism and clearance
Metabolite ID

In-vitro toxicity testing applications:
Hepatotoxicity (reactive metabolites, lipidosis, choleastasis etc)

Virology (HBV, HCV…)
Infection with hepatitis and other viruses for replication


Innovative Modes of Culture
BioArtificial Livers 
Transgenic liver humanized animals with HepaRG™ cells
…. an ever-expanding range of applications!

HepaRG™ Key Advantages

- The capacity to produce early hepatic progenitor cells as well as completely mature human hepatocytes.
- A morphology and metabolic capacity similar to human primary hepatocytes.
- The first human hepatoma cell line to express main CYPs, functional nuclear receptors and major sinusoidal and canalicular hepatic drug transporters.
- Overcomes limited availability of liver tissue.
- Excellent batch-batch reproducibility and sub-cultures.
- CYP activities and expression of canalicular drug transporters can be increased by classical inducers.
- A robust cell-based system to evaluate metabolism of test compounds.
- A suitable model for high throughput screening of chemicals.
- A unique and exclusive model for in vitro pharmacological and toxicological studies.
- Usable and stable for at least 4 weeks once differentiation is achieved, ideal for long-term studies.
- Can support the complete replicative cycle of HBV.


In-vitro ADME Applications

Drug metabolism and clearance
HepaRG™ cells be classified as an “average human hepatocyte population” and express multiple functional Phase 1 and 2 drug metabolising enzyme (DME) activities comparable to levels in cultures of primary human hepatocytes. This makes them a much better model for metabolism and clearance studies than any other cell line currently available. The prediction of in-vivo intrinsic clearance of a panel of drugs has been shown to be mostly within 2fold using HepaRG™ cells. Moreover, the sustained presence of DMEs over weeks makes them ideal for measuring the clearance of low clearance compounds.

Metabolite ID
HepaRG™ cells have a full complement of DMEs, cofactors and transporter proteins and the profile of DME activities and expression provides a good representation of that in primary human hepatocytes. Together with the high stability of the enzymes, this makes these cells ideal for producing relevant drug metabolites in sufficient quantities to enable metabolite identification.

DME induction assays
HepaRG™ cells express functional transcription factors involved in the regulation of DMEs. These cells are responsive to prototypical inducers of CYP1A1/2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP3A4 and can be used to predict in-vivo CYP3A4 induction of test compounds by constructing a calibration curve using known potent and mild inducers of this CYP (“AUC/F2” or “RIS”). The protocol for CYP induction using HepaRG™ cells has been accepted by ESAC and an OECD guideline for use of HepaRG™ cells in induction studies is in preparation.

DME inhibition assays
The high levels of DMEs in HepaRG™ cells makes them ideal for inhibition screening studies. The IC50 values of a panel of CYP inhibitors using HepaRG™ cells correlated very well with those determined in primary human hepatocytes. Time-dependent inhibition can also be investigated using these cells.

Functional transporter assays
HepaRG™ cells express a number of uptake and efflux drug transporters and form tight junctions and bile canaliculi, making them ideal for uptake and biliary secretion studies.

In-vitro toxicity testing applications

Hepatotoxicity Screening and Mechanistic Testing Applications

Carbohydrate and lipid metabolism
HepaRG™ cells display glucose and insulin responsiveness and can be used to investigate carbohydrate homeostasis and lipogenesis.

Reactive metabolites
The stable and high levels of DMEs in HepaRG™ cells make them ideal for long-term and repeated dose toxicity studies in which a compound may be bioactivated and/or cause latent effects over days or weeks.

Steatosis
The accumulation triglycerides in HepaRG™ cells is predictive of the in-vivo steatosis reported in patients chronically treated with steatogenic drugs. Lipid accumulation directly correlates with levels of mRNA encoding for lipid synthesis markers, thus this toxicity can be monitored visually and at the gene level.

Phospholipidosis
HepaRG™ cells can be used to discriminate between drugs which cause steatosis (after 24 h) and phospholipidosis (evident after 2 weeks) e.g. amiodarone; and those which cause steatosis only e.g. tetracycline.

Cholestasis
Cholestatic drugs (via direct inhibition or down-regulation) can be identified by measuring their effects on bile acid efflux in HepaRG™ cells.

Genotoxicity and carcinogenicity
Since HepaRG™ cells have a good metabolic capacity and proliferative capacity, they can be implemented into the micronucleus test (MNT), as recommended by ECVAM and IWGT workshops. Whole genome expression analysis of HepaRG™ cells can discriminate between genotoxic and non-genotoxic compounds and provide information on compound- and time-dependent effects on cell cycle and apoptosis signalling pathways.

Mitochondrial toxicology
Apoptosis
Inflammation

Virology and Infection

HBV, HCV, HEV
HepaRG™ cells support viral replication can be used to model viral infection and provide insight into the processes involved in virus receptor binding, uptake, and membrane fusion

Malaria

Innovative Modes of Culture

BioArtificial Livers (BALs)
HepaRG™ cells can be cultured in 3D bioreactors to generate an in-vivo-like assembly of highly polarised cells with high and sustained CYP activities. This makes them a promising cell type for use as liver assist devices or BALs.

Transgenic liver humanized animals with HepaRG™ cells
HepaRG™ cells can be transplanted into chimeric mice to repopulate their livers and express human primary hepatocyte–specific genes and proteins. Such a HepaRG™-mouse combination represents a promising model for in-vivo studies of drug metabolism and hepatitis virus infections.

Spheroids
Micropatterns
Dynamic fluidic models
Co-culture models
Stem cell cultures



Caltag Medsystems are the exclusive UK and Ireland distributor for Biopredic products. For further information about HepaRG or any of their products please call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk

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